Abstract − Analytical Sciences, 23(3), 271 (2007).
A Peptide Microarray for the Detection of Protein Kinase Activity in Cell Lysate
Syuhei SHIGAKI,*1 Takayuki YAMAJI,*2 Xiaoming HAN,*2 Go YAMANOUCHI,*2 Tatsuhiko SONODA,*3 Osamu OKITSU,*4 Takeshi MORI,*1,*5 Takuro NIIDOME,*1,*5 and Yoshiki KATAYAMA*1,*5
*1 Department of Applied Chemistry, Faculty of Engineering, Kyushu University, 744 Motooka, Nishi, Fukuoka 819-0395, Japan
*2 Graduate School of Systems Life Sciences, Kyushu University, 744 Motooka, Nishi, Fukuoka 819-0395, Japan
*3 Department of Materials Science and Chemical Engineering, Kitakyushu National College of Technology, 5-20-1 Shii, Kokuraminami, Kitakyushu 802-0985, Japan
*4 Applied Genomics, Molecular Medicine Research Laboratories, Astellas Pharmaceuticals Inc., 5-2-3 Tokoudai, Tsukuba 330-2698, Japan
*5 Center for Future Chemistry, Kyushu University, 744 Motooka, Nishi, Fukuoka 819-0395, Japan
*2 Graduate School of Systems Life Sciences, Kyushu University, 744 Motooka, Nishi, Fukuoka 819-0395, Japan
*3 Department of Materials Science and Chemical Engineering, Kitakyushu National College of Technology, 5-20-1 Shii, Kokuraminami, Kitakyushu 802-0985, Japan
*4 Applied Genomics, Molecular Medicine Research Laboratories, Astellas Pharmaceuticals Inc., 5-2-3 Tokoudai, Tsukuba 330-2698, Japan
*5 Center for Future Chemistry, Kyushu University, 744 Motooka, Nishi, Fukuoka 819-0395, Japan
DNA microarray enables the analysis of DNA or mRNA expression levels, but it has not been possible to completely understand life using obtained information. Consequently, protein or peptide arrays have attracted much interest. Since the development of a practical protein microarray is still far away in light of handling difficulties, the peptide microarray is a promising tool for analyzing protein functions. We have developed a peptide microarray to detect protein kinase activity in cell lysate. All substrate peptides for kinases were immobilized chemoselectively on amino-coated glass slides. After phosphorylation of the immobilized peptides, phosphorylation was detected by fluorescence imaging. We detected the protein kinase activities, including that in cell lysate, in response to drug stimulation. Therefore, this peptide microarray would be useful for a high-throughput kinase assay of intracellular signals and would be applicable to drug screening.
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